Discovery of novel therapies for patients with CF not eligible for registered modulator treatments, assisted by novel microfluidic techniques for combinatorial, high-throughput organoid screens

Perspectives for patients with cystic fibrosis (CF) have dramatically changed oPerspectives for patients with cystic fibrosis (CF) have dramatically changed over the last few years, with the advent of CFTR modulators, a new treatment claver the last few years, with the advent of CFTR modulators, a new treatment class comprising several mutation-specific drugs. Recent drug development efforts ss comprising several mutation-specific drugs. Recent drug development efforts are directed to the most frequent CFTR mutation (F508del) and a handful of lessare directed to the most frequent CFTR mutation (F508del) and a handful of less frequent mutations, excluding around 10% of patients with only rare mutations, frequent mutations, excluding around 10% of patients with only rare mutations, whom depend on academic research innovation and public-private partnerships to whom depend on academic research innovation and public-private partnerships to provide efficient treatments.Intestinal organoids allow a personalized medic provide efficient treatments.Intestinal organoids allow a personalized medicine approach to identify drug combinations or gene editing therapies that rescuine approach to identify drug combinations or gene editing therapies that rescue CFTR function (measured by CFTR-dependent organoid swelling). However, this te CFTR function (measured by CFTR-dependent organoid swelling). However, this technique is labor and cost intensive and innovations increasing the throughput echnique is labor and cost intensive and innovations increasing the throughput paralleled by a reduction in sample size and reagent volumes, would be of greatparalleled by a reduction in sample size and reagent volumes, would be of great economical and societal value.Here, we will investigate, on biobanked rectal economical and societal value.Here, we will investigate, on biobanked rectal organoids of ~70 CF patients with rare mutations ineligible for licensed drugs organoids of ~70 CF patients with rare mutations ineligible for licensed drugs:(1) Novel combinations of CFTR modulators, approved or in late development p:(1) Novel combinations of CFTR modulators, approved or in late development phase, in their ability to restore CFTR function(2) For mutations not rescuablhase, in their ability to restore CFTR function(2) For mutations not rescuable by compounds, we will develop tailored CRISPR-Cas-based gene editing strategie by compounds, we will develop tailored CRISPR-Cas-based gene editing strategies.To scale up the process, we will develop a novel microfluidic High-Throughes.To scale up the process, we will develop a novel microfluidic High-Throughput Screening (HTS) platform, with increased test capacity on smaller organoid put Screening (HTS) platform, with increased test capacity on smaller organoid samples, opening up avenues for rapid and ex vivo HTS for drug and gene editingsamples, opening up avenues for rapid and ex vivo HTS for drug and gene editing therapies in a personalized medicine approach. therapies in a personalized medicine approach.

Keywords:Gene editing, Microfluidics, Cystic Fibrosis, Crispr-Cas, CFTR modulators

Disciplines:Respiratory medicine, Microfluidics/flow chemistry